title: Optimal detection of sentinel lymph node metastases by intraoperative radioactive threshold and molecular analysis in patients with melanoma.
abstract: The aim of this study was to optimize a protocol for radioguided biopsy of the sentinel lymph node (SLN) in patients with melanoma. The protocol was based on a combination of ex vivo counting of the nodes detected intraoperatively and analysis of the harvested nodes by hematoxylin and eosin staining plus immunohistochemistry (conventional histopathology [PATH]) and by molecular biology (reverse-transcriptase polymerase chain reaction [RT-PCR]).
  
  
  METHODS
  A total of 124 patients with primary clinical stage I-II (according to the American Joint Committee on Cancer) cutaneous melanoma underwent successful radioguided SLN biopsy. SLNs harvested for analysis included any additional nodes whose ex vivo counting rate exceeded 20% of the hottest node. All removed SLNs were examined by conventional PATH and with RT-PCR analysis for the expression of messenger RNA for tyrosinase and the melanoma antigens recognized by T cells. Complete lymph node dissection (CLND) was performed only in the case of SLN metastasis detected by PATH. Different combinations of the intraoperative parameters (only the hottest node and all nodes harvested) and of analysis (PATH and RT-PCR) were tested as predictors of clinical outcome on the basis of long-term follow-up (12-81 mo; median, 55 mo).
  
  
  RESULTS
  A total of 197 SLNs were harvested, 41 of which harbored metastasis as detected by RT-PCR analysis; PATH detected metastasis in only 24 of 41 metastatic SLNs. In 5 of 41 instances, metastasis was not in the hottest SLN. The main factor determining correct classification of the SLN status was RT-PCR, which significantly improved detection of metastasis, even if applied only to the hottest node (P < 0.0001 vs. PATH analysis of either the hottest SLN or all nodes above the 20% threshold). Metastatic disease recurred locally in 5 patients who had not undergone CLND; RT-PCR analysis showed metastasis in 4 of these patients. The false-negative rate of SLN biopsy progressively decreased when applying PATH only to the hottest node (32.1%), additional RT-PCR to the hottest node (21.4%), PATH to all nodes (17.9%), and RT-PCR to all nodes (3.6%, P = 0.015 vs. PATH analysis of only the hottest SLN).
  
  
  CONCLUSIONS
  On the basis of long-term follow-up (the gold standard for final clinical outcome of SLN biopsy), both 20% threshold and RT-PCR analysis should be applied for optimal detection of nodal metastases in patients with melanoma.
authors.affiliation: Regional Center of Nuclear Medicine, University of Pisa Medical School, Pisa, Italy. g.manca@med.unipi.it
authors.forename: Gianpiero
authors.initials: G
authors.lastname: Manca
authors.forename: Antonella
authors.initials: A
authors.lastname: Romanini
authors.forename: Daniela
authors.initials: D
authors.lastname: Pellegrino
authors.forename: Elisa
authors.initials: E
authors.lastname: Borsò
authors.forename: Marianna
authors.initials: M
authors.lastname: Rondini
authors.forename: Cinzia
authors.initials: C
authors.lastname: Orlandini
authors.forename: Vanna
authors.initials: V
authors.lastname: Zucchi
authors.forename: Francesco
authors.initials: F
authors.lastname: Pasqualetti
authors.forename: Giuliano
authors.initials: G
authors.lastname: Mariani
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D004801:Eosine Yellowish-(YS); D005188:False Negative Reactions; D005260:Female; D005500:Follow-Up Studies; D006416:Hematoxylin; D006801:Humans; D007150:Immunohistochemistry; D007432:Intraoperative Period; D008197:Lymph Node Excision; D008198:Lymph Nodes / Q000378:metabolism* / Q000473:pathology / Q000601:surgery*; D008207:Lymphatic Metastasis; D008297:Male; D008545:Melanoma / Q000473:pathology*; D008875:Middle Aged; D012313:RNA / Q000235:genetics / Q000378:metabolism; D011851:Radioactivity; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012306:Risk; D021701:Sentinel Lymph Node Biopsy / Q000379:methods*; D013194:Staining and Labeling; D013696:Temperature; D016896:Treatment Outcome; D055815:Young Adult
chemical_list: D012313:RNA; D004801:Eosine Yellowish-(YS); D006416:Hematoxylin